DNA提取与纯化

1.加入solution.III，经10分钟离心后细菌沉淀怎么不结实，有的漂浮在液面，有的贴在离心管壁上，一摇晃即破碎脱落下来？细菌的用量太少，导致产生的沉淀主要是盐分的沉淀，因为缺少变性的细菌蛋白和细菌基因组DNA 的缠绕，沉淀就显得不结实。解决方法：将细菌的用量增加。 2.加入soln.III，经10分钟离心后细菌沉淀怎么不结实，呈大块的水泡状，上清较少？（1）使用了过多的细菌，导 ...

近10年来，现代分子生物学技术越来越广泛地被用于人类疾病研究的诸领域，为了解病理状态下DNA的变化积累了新资料。目前认为，人类并非人们想像的那样稳定，诸如基因重排、扩增、缺失，突变和DNA甲基化类型改变等时有发生，这些改变对于基因表达和调控，以及疾病过程的发展与转归等方面均具有重要意义。 　　医院病理科档案中积存的大量石蜡包埋组织，是一个可靠的分子生物学研究的材料来源。在石蜡切片上进行原位杂交或原 ...

Jacks Lab Center for cancer researchMIT 1. Each tail should be in a clean eppendorf tube. 2. Add 500µl of tail lysis buffer containing Proteinase K (PK) to each tube. 3. Incubate tai ...

1. A ...

一.实验目的 1.掌握DNA 指纹图谱技术的概念、原理和基本操作过程 2.学习DNA 的限制性酶切的基本技术 3.掌握琼脂糖凝胶电泳的基本操作技术，学习利用琼脂糖凝胶电泳测定DNA 片段的长度，并能对实验结果进行分析。 二.实验原理 1984 年英国莱斯特大学的遗传学家 Jefferys 及其合作者首次将分离的人源小卫星DNA 用作基因探针，同人体核DNA 的酶切片段杂交，获得了 ...

DNA Fragment Purification from Agarose or Acrylamide For fragments from 200 bp to 10 kb the agarose purification is ideal.For smaller fragments (20 bp to 400 bp)the acrylamide purification is preferre ...

1.grow plants in trays of 96 and leave two spots open (for the PCR controls) 2.harvest 1 to 2 young and green leaves (1cm2 /plant at rosette stage if possible). Use 96 well plates (1 or 2ml E&K p ...

Pick colonies into 0.5ml of SD-Leu (or other appropriate SD medium) Vortex for 1min Leave to grow O/N for 18-24h at 30℃ 230-250rpm (best in 5ml bijou) Spin yeast culture at 13000rpm for 5 min (microfu ...

适合从动物细胞、动物组织或酵母菌中同步纯化总RNA 和基因组DNA纯化的总RNA ，适合用于Northern Blot、RT-PCR、体外翻译、Primer Extension、S1 核酸酶作图、RNase 保护测定、制备mRNA 和构建cDNA 文库等各种RNA 分子生物学实验。 纯化的基因组DNA 适用于PCR、Southern Blot、RAPD、AFLP、RFLP 等分子生物学实验。 一、 ...

David Harry Institute of Forest Genetics USDA Forest Service Pacific Southwest Research Station August 26 1993 Background : There are many published methods for mini-preps of DNA from lambda phage cl ...

1.Obtain 65-100 µl of blood by retro-orbital bleed with a heparinized microcapillary tube.Expel blood immediately into a 1.5 ml microfuge tube containing 20 µl of 10 mM EDTA.Mix immediatel ...

第一节 概 述 DNA的提取通常用于构建文库、Southern杂交(包括RFLP)及PCR分离基因等。利用DNA较长的特性，可以将其与细胞器或质粒等小分子DNA分离。加入一定量的异丙醇或乙醇，的大分子DNA即沉淀形成纤维状絮团飘浮其中，可用玻棒将其取出，而小分子DNA则只形成颗粒状沉淀附于壁上及底部，从而达到提取的目的。在提取过程中，染色体会发生机械断裂，产生大小不同的片段，因此分离DNA时应尽量 ...

1 目的 ：DNA 抽提标准化操作规范适用于HLA试验中的DNA 抽提操作。 2 适用范围 ：DNA 抽提实验室 3 依据 ： DNA 抽提标准化操作规范 4 引用文件 ：德国BAG公司（以下简称BAG）《Instructions for use EXTRA-GENE I》 5 程序 5.1试剂准备 5.1.1 BAG® EXTRA-GENE I 该试剂盒包括3部分：Solution 1 ...

1.Prepare or obtain Buffered phenolpH 8.Add 2 crystals of 8-Hydroxyquinoline to prevent oxidation.This also identifies the organic phase as yellow-colored. 2.Combine DNA sample with an equal volume of ...

1.Separate DNA fragments in an agarose gel cast with 0.5 mg/ml Ethidium bromide.Locate bands with a hand-held long-wave UV lamp. 2.Slice the gel with a razor blade above and below the bands of interes ...

A protocol / method / schedule /procedure for extraction / isolation of both DNA and RNA from the same material typically plant leaf / leaves (See also 1)Take one medium sized leaf or half a large lea ...

This procedure was originally developed for Listeria monocytogenes but has worked well with other Gram+ bacteria we've tried. 1.Pellet cells from 10 ml overnight cultures in BHI or LB and wash in 5 ml ...

1.Pour a vertical acrylamide gel using TEA buffer.A 4 % non denaturing gel is correct for most applications. 2.Run out DNA fragments.For fragments greater than 500 bprun the xylene cyanol dye to the b ...

This is a modification of the procedure in Short Protocols (1-411-45) 1.Prepare a 50 ml liquid lysate: A.mix 2xlO8 E.coli cells with 100μl phage (from one picked plaque in 500μl SM)and 100μl ...

(adapted from Bruce A.RoeDepartment of Chemistry and BiochemistryThe University of OklahomaNormanOklahoma 73019 broe@ou.edu) Phenol extraction is a common technique used to purify a DNA sample (1).Typ ...